miRNA profiling in human precision-cut lung slices (PCLS)

Transcriptome analysis revealed miRNA profiles from PCLS of tumor-free tissue comparable to normal lung tissue Suitability of human PCLS as a model for lung-specific miRNA investigations by RTqPCR and microarray analyses miRNA profiles from human PCLS of tumor-free tissues in n = 10 donors were obtained by GeneChip™ miRNA 4.0 Arrays. To determine their use for gene expression analysis, the miRNA profiles (tumor-free tissues) were compared to miRNA profiles (also obtained by GeneChip™ miRNA 4.0 Arrays) of n = 9 human PCLS treated with low dose (LD) 0.1 mM; medium dose (MD) 0.3 mM; and high dose (HD) 1 mM of methyl acrylate. To determine whether the expression levels of certain lung-disease associated miRNAs expressed in the human PCLS of tumor-free tissue comparable to miRNA expression levels in ‘healthy’ lung, we searched for published miRNA data from human lungs. CEL Files from GSE81203 obtained by GeneChip™ miRNA 3.0 Array were downloaded from GEO and were re-analyzed TAC 4.0, using Affymetrix default settings and RMA normalization. Lung disease-related miRNAs with average signal (log2) intensity > 6 in the human PCLS of tumor-free tissues were compared manually to corresponding average signal (log2) intensity in the ‘healthy’ lung samples (GSE81203).